{"title":"Development of a highly specific LAMP assay for detection of Sarcocystis tenella and Sarcocystis gigantea in sheep","authors":"Yajie Chen, Jing Peng, Zifu Zhu, Wen Zhang, Lifang Wang, Jianhai Xu, Qun Liu, Jing Liu","doi":"10.1007/s00436-024-08349-0","DOIUrl":null,"url":null,"abstract":"<p><i>Sarcocystis</i> infection in sheep has caused significant economic losses in the livestock industry, and the genetic similarity among <i>Sarcocystis</i> species highlights the need for precise diagnostic methods in sheep. This study developed a loop-mediated isothermal amplification (LAMP) method targeting <i>COX-1</i> and <i>28S rRNA</i> genes to detect <i>Sarcocystis tenella</i> and <i>Sarcocystis gigantea</i>, respectively. The LAMP method exhibited high specificity, selectively amplifying target DNA sequences without cross-reactivity with closely related protozoa, such as <i>Toxoplasma gondii</i> and <i>Neospora caninum</i>. Detection limits were determined as 3 × 10<sup>5</sup> copies/L for <i>S. tenella</i> and 6 × 10<sup>4</sup> copies/L for <i>S. gigantea</i>, enabling sensitive identification of low-level infections. Comparative analysis with conventional PCR on sheep cardiac tissues demonstrated a higher LAMP detection rate (80.0% vs 66.7%). In conclusion, the LAMP method offers superior sensitivity to conventional PCR, allows visual confirmation of results, and provides a rapid diagnostic tool for identifying <i>S. tenella</i> and <i>S. gigantea</i> infection in sheep. However, due to the limitation of sample availability, we were unable to assess all <i>Sarcocystis</i> species that use sheep as intermediate hosts, which warrants further research.</p>","PeriodicalId":19968,"journal":{"name":"Parasitology Research","volume":null,"pages":null},"PeriodicalIF":1.8000,"publicationDate":"2024-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Parasitology Research","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1007/s00436-024-08349-0","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"PARASITOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
Sarcocystis infection in sheep has caused significant economic losses in the livestock industry, and the genetic similarity among Sarcocystis species highlights the need for precise diagnostic methods in sheep. This study developed a loop-mediated isothermal amplification (LAMP) method targeting COX-1 and 28S rRNA genes to detect Sarcocystis tenella and Sarcocystis gigantea, respectively. The LAMP method exhibited high specificity, selectively amplifying target DNA sequences without cross-reactivity with closely related protozoa, such as Toxoplasma gondii and Neospora caninum. Detection limits were determined as 3 × 105 copies/L for S. tenella and 6 × 104 copies/L for S. gigantea, enabling sensitive identification of low-level infections. Comparative analysis with conventional PCR on sheep cardiac tissues demonstrated a higher LAMP detection rate (80.0% vs 66.7%). In conclusion, the LAMP method offers superior sensitivity to conventional PCR, allows visual confirmation of results, and provides a rapid diagnostic tool for identifying S. tenella and S. gigantea infection in sheep. However, due to the limitation of sample availability, we were unable to assess all Sarcocystis species that use sheep as intermediate hosts, which warrants further research.
期刊介绍:
The journal Parasitology Research covers the latest developments in parasitology across a variety of disciplines, including biology, medicine and veterinary medicine. Among many topics discussed are chemotherapy and control of parasitic disease, and the relationship of host and parasite.
Other coverage includes: Protozoology, Helminthology, Entomology; Morphology (incl. Pathomorphology, Ultrastructure); Biochemistry, Physiology including Pathophysiology;
Parasite-Host-Relationships including Immunology and Host Specificity; life history, ecology and epidemiology; and Diagnosis, Chemotherapy and Control of Parasitic Diseases.